{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T21:31:45Z","timestamp":1768512705875,"version":"3.49.0"},"posted":{"date-parts":[[2024,6,10]]},"group-title":"Biochemistry","reference-count":52,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,6,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Purinergic P2X3 receptors form trimeric cation-gated channels, which are activated by extracellular ATP. P2X3 plays a crucial role in chronic cough and affects over 10% of the population. Despite considerable efforts to develop drugs targeting P2X3, the highly conserved structure within the P2X receptor family presents obstacles for achieving selectivity. Camlipixant, a potent and selective P2X3 antagonist, is currently in phase III clinical trials. However, the mechanisms underlying receptor desensitization, ion permeation, principles governing antagonism, and the structure of P2X3 when bound to camlipixant remain elusive. In this study, we established a stable cell line expressing homotrimeric P2X3 and utilized a peptide scaffold to purify the complex and determine its structure using cryo-electron microscopy (cryo-EM). P2X3 binds to camlipixant at a previously unidentified drug-binding site and functions as an allosteric inhibitor. Structure-activity studies combined with modeling and simulations have shed light on the mechanisms underlying the selective targeting and inhibition of P2X3 by camlipixant, distinguishing it from other members of the P2X receptor family.<\/jats:p>","DOI":"10.1101\/2024.06.10.598171","type":"posted-content","created":{"date-parts":[[2024,6,10]],"date-time":"2024-06-10T17:40:11Z","timestamp":1718041211000},"source":"Crossref","is-referenced-by-count":1,"title":["A Second Drug Binding Site in P2X3"],"prefix":"10.64898","author":[{"given":"Trung","family":"Thach","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"KanagaVijayan","family":"Dhanabalan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Prajwal Prabhakarrao","family":"Nandekar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seth","family":"Stauffer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iring","family":"Heisler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah","family":"Alvarado","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan","family":"Snyder","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6709-190X","authenticated-orcid":false,"given":"Ramaswamy","family":"Subramanian","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024061723500737000_2024.06.10.598171v2.1","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1016\/S0140-6736(14)61255-1","article-title":"P2X3 receptor antagonist (AF-219) in refractory chronic cough: a randomised, double-blind, placebo-controlled phase 2 study","volume":"385","year":"2015","journal-title":"The Lancet"},{"key":"2024061723500737000_2024.06.10.598171v2.2","doi-asserted-by":"publisher","DOI":"10.1107\/S2059798318006551"},{"key":"2024061723500737000_2024.06.10.598171v2.3","doi-asserted-by":"crossref","first-page":"e53530","DOI":"10.7554\/eLife.53530","article-title":"New approach for membrane protein reconstitution into peptidiscs and basis for their adaptability to different proteins","volume":"9","year":"2020","journal-title":"eLife"},{"key":"2024061723500737000_2024.06.10.598171v2.4","doi-asserted-by":"publisher","DOI":"10.1378\/chest.129.1_suppl.238S"},{"key":"2024061723500737000_2024.06.10.598171v2.5","doi-asserted-by":"publisher","DOI":"10.1378\/chest.14-1481"},{"key":"2024061723500737000_2024.06.10.598171v2.6","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.34085"},{"key":"2024061723500737000_2024.06.10.598171v2.7","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.34085"},{"key":"2024061723500737000_2024.06.10.598171v2.8","doi-asserted-by":"publisher","DOI":"10.1007\/s10858-015-9969-9"},{"key":"2024061723500737000_2024.06.10.598171v2.9","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.pupt.2011.01.012","article-title":"Chronic \u2018cough hypersensitivity syndrome\u2019: A more precise label for chronic cough","volume":"24","year":"2011","journal-title":"Pulmonary Pharmacology & Therapeutics"},{"key":"2024061723500737000_2024.06.10.598171v2.10","doi-asserted-by":"crossref","unstructured":"Corriden R & Insel PA (2010) Basal Release of ATP: An Autocrine-Paracrine Mechanism for Cell Regulation. 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